A Functional Interface at the rDNA Connects rRNA Synthesis, Pre-rRNA Processing and Nucleolar Surveillance in Budding Yeast

• Published in: PloS one Vol. 6; no. 9; p. e24962
• Main Authors: Leporé, Nathalie, Lafontaine, Denis L. J.
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 19.09.2011; Public Library of Science (PLoS)
• Subjects: Aberration; Binding; Biology; Blotting, Northern; Blotting, Western; Cell Nucleolus - genetics; Cell Nucleolus - metabolism; Chromatin Immunoprecipitation; Chromosomal Proteins, Non-Histone - genetics; Chromosomal Proteins, Non-Histone - metabolism; DNA, Ribosomal - metabolism; DNA-directed RNA polymerase; Elongation; Kinases; Metabolism; Morphology; Mutation; Mutation - genetics; Nuclear Proteins - genetics; Nuclear Proteins - metabolism; Nucleoli; Nucleotide sequence; Phosphorylation; Polyadenylation; Protein binding; Proteins; Quality control; Real-Time Polymerase Chain Reaction; Recruitment; Ribonucleic acid; Ribosomal RNA; Ribosomes; RNA; RNA polymerase II; RNA Polymerase II - genetics; RNA Polymerase II - metabolism; RNA Precursors - genetics; RNA Precursors - metabolism; RNA processing; RNA Processing, Post-Transcriptional; RNA synthesis; RNA, Messenger - genetics; RNA-Binding Proteins - genetics; RNA-Binding Proteins - metabolism; rRNA; Saccharomyces cerevisiae Proteins - genetics; Saccharomyces cerevisiae Proteins - metabolism; Saccharomycetales - genetics; Saccharomycetales - growth & development; Saccharomycetales - metabolism; Surveillance; Synthesis; Transcription (Genetics); Transcriptional Elongation Factors - genetics; Transcriptional Elongation Factors - metabolism; Yeast; Belgium
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0024962

Ribogenesis is a multistep error-prone process that is actively monitored by quality control mechanisms. How ribosomal RNA synthesis, pre-rRNA processing and nucleolar surveillance are integrated is unclear. Nor is it understood how defective ribosomes are recognized. We report in budding yeast that, in vivo, the interaction between the transcription elongation factor Spt5 and Rpa190, the largest subunit of RNA polymerase (Pol) I, requires the Spt5 C-terminal region (CTR), a conserved and highly repetitive domain that is reminiscent of the RNA Pol II C-terminal domain (CTD). We show that this sequence is also required for the interaction between Spt5 and Nrd1, an RNA specific binding protein, and an exosome cofactor. Both the Spt4-Spt5, and the Nrd1-Nab3 complexes interact functionally with Rrp6, and colocalize at the rDNA. Mutations in the RNA binding domain of Nrd1, but not in its RNA Pol II CTD-interacting domain, and mutations in the RRM of Nab3 led to the accumulation of normal and aberrant polyadenylated pre-rRNAs. Altogether these results indicate that Nrd1-Nab3 contributes to recruiting the nucleolar surveillance to elongating polymerases to survey nascent rRNA transcripts.